雷公藤内酯醇减少放射性肺纤维化中肌成纤维细胞活化与抑制TGF-β1/ERK/Smad3通路相关  

TPL's suppression on activation of myofibroblasts in radiation induced lung fibrosis related to its inhibition on TGF-β1/ERK/Smad3 pathway

作　　者：张彦伟[1] 张志强[1,2,3] 吴丽贤[1,2,3] 张鲁榕[4,5] 陈纯[1,2,3] ZHANG Yan-wei ZHANG Zhi-qiang WU Li-xian ZHANG Lu-rong CHEN Chun(Dept of Pharmacology Institute of Materia Medica Fujian Key Laboratory of Natural Medicine Pharmacology,Fujian Medical University, Fuzhou 350122,China the First Affiliated Hospital of Fujian Medical University,Fuzhou 350004,China Key Laboratory of Radiation Biology, Fujian Medical University, Fuzhou 350004,China)

机构地区：[1]福建医科大学药学院药理学系,福建福州350122 [2]福建医科大学新药研究所,福建福州350122 [3]福建医科大学福建省天然药物药理学重点实验室,福建福州350122 [4]福建医科大学附属第一医院,福建福州350004 [5]福建医科大学放射生物学重点实验室,福建福州350004

出　　处：《中国药理学通报》2017年第5期630-636,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81473264);福建省卫生厅中医药科研项目(No wzze201308);福建省自然科学基金面上项目(No 2014J01336)

摘　　要：目的通过体内外实验,观察雷公藤内酯醇(TPL)减少放射性肺纤维化中肌成纤维细胞(MFBs)活化与TGF-β1/ERK/Smad3通路的相关性。方法以TGF-β1刺激成纤维细胞建立体外MFBs活化模型,C57BL/6小鼠胸部照射形成体内MFBs活化、放射性肺纤维化模型。MFBs活化状态通过检测小鼠成纤维细胞中α-SMA(RT-PCR、Western blot方法)和ColⅠ(RT-PCR、ELISA方法)的表达,通路活性采用Western blot法检测p-ERK、p-Smad3(Ser208)、p-Smad3(Ser423)的水平。ERK siRNA及Smad3 siRNA观察ERK、Smad3在MFBs活化中的地位。结果 TGF-β1激活p-ERK/p-Smad3(Ser208)及p-Smad3(Ser423),诱导成纤维细胞表达α-SMA,活化为MFBs,合成Col I明显增多;使用ERK siRNA及Smad3 siRNA确定了ERK及Smad3均参与α-SMA、ColⅠ的表达,其中ERK可能通过磷酸化Smad3连接区(Ser208)起相关作用。小鼠胸部照射可致肺组织中p-ERK、p-Smad3(Ser208)、p-Smad3(Ser423)的水平上调,α-SMA表达增加,显示多量MFBs活化。TPL对体内和体外实验中ERK、Smad3(Ser208)、Smad3(Ser423)的磷酸化活化均有明显抑制作用,可明显下调α-SMA表达及ColⅠ合成,减少MFBs的活化。结论 TPL可通过抑制TGF-β1/ERK/Smad3通路,减少肺部照射后MFBs活化,从而抑制放射性肺纤维化进展。Aim To observe the correlation between the TPL＇s suppression on myofibrolbasts（MFBs）activation and TGF-β1/ERK/Smad3 pathway by performing in vivo and in vitro experiments.Methods In vitro model of MFBs activation was set up by stimulating fibroblasts with TGF-β1, and in vivo model of MFBs activation in radiated lung tissue was built by thoracic radiation on C57BL/6 mice.MFBs activation was analyzed by detecting the expression of α-SMA（using RT-PCR and Western blot）and Col Ⅰ（using RT-PCR and ELISA methods）.The levels of p-ERK, p-Smad3（Ser208）and p-Smad3（Ser423）were measured by Western blot.ERK siRNA and Smad3 siRNA were used to observe the status of ERK and Smad3 in MFBs activation.Results TGF-β1 activated p-ERK/p-Smad3（Ser208）and p-Smad3（Ser423）, increased the expression of α-SMA and synthesis of Col Ⅰ, which indicated MFBs activation.siRNA knockdown assay showed that both ERK and Smad3 were involved in regulating the levels of α-SMA and Col Ⅰ, and ERK influenced MFBs transformation possibly through its phosphorylation of Smad3（Ser208）.TPL treatment inhibited the phosphorylation activation of ERK, Smad3（Ser208）, Smad3（Ser423）in vitro and in vivo, therefore significantly reduced the level of α-SMA and Col Ⅰ, and the number of activated MFBs was decreased.Conclusion TPL mitigates radiation-induced pulmonary fibrosis by inhibiting the activation of MFBs, which is partly through suppressing TGF-β1/ERK/Smad3 pathway.

关 键 词：雷公藤内酯醇 放射性肺纤维化 肌成纤维细胞 Α-平滑肌肌动蛋白 Ⅰ型胶原蛋白 转化生长因子β1 ERK SMAD3 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R563.102.2[医药卫生—基础医学]